The complete mitochondrial genome of Vibrissina turrita (Meigen, 1824) (Diptera, Tachinidae)

Abstract In this study, the mitogenome of Vibrissina turrita (Meigen, 1824) (Diptera, Tachinidae) was sequenced based on the next-generation sequencing approach and analyzed here for the first time. The 17,387 bp genome has a high A + T content and consists of 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes, and one noncoding control region. The phylogenetic analysis results support that Exoristinae is monophyletic and V. turrita belongs to the subfamily. This study reveals the systematic classification status of V. turrita and will enrich the genetic data on Tachinidae.


Introduction
Exoristinae is the largest subfamily in the family Tachinidae (Diptera), which has 3,687 described species worldwide (O'Hara et al. 2020).As a parasitoid, it is a significant natural enemy in natural and managed terrestrial ecosystems, particularly in forests (Yan et al. 2021).It has a wide range of hosts, mainly parasitizing on larvae of Lepidoptera, and is also a natural enemy of certain Orthoptera, Hymenoptera, and other insect orders (Stireman et al. 2006).Vibrissina Rondani belongs to the Blondeliini, with a total of 43 species, of which three have been recorded in China (O'Hara et al. 2020).Mitochondrial genome has been widely used in phylogenetic studies or molecular level species identification of Diptera insects, e.g.Yan et al. (2021) and Zhou et al. (2024).But only a few complete mitochondrial genomes of the Tachinidae have been published.In this study, we sequenced the mitogenome data of V. turrita (Meigen, 1824) and annotated it as the first mitogenome of genus Vibrissina, which will contribute to gaining an understanding of the phylogenetic relationships of V. turrita and future genetic research in the family.
According to the manufacturer's protocol, a QIAamp Micro DNA Kit (Qiagen, Hilden, Germany) was used to extract DNA from the female adult's muscle tissues of the thorax (the voucher number for the sample is SYNU20210808).Genomic DNA was randomly fragmented by Covaris (Covaris, LLC, Woburn, MA), followed by fragments selection by Agencourt AMPure XP-Medium Kit (Beckman, Brea, CA) to an average size of 200-400 bp.Selected fragments were end repaired and 3 0 adenylated, then the adaptors were ligated to the ends of these 3 0 adenylated fragments.Finally, the single strand circle DNA was formatted as the final library.DNBSEQ-T7 (MGI, Shenzhen, China) was used to perform paired-end sequencing with a read length of 150 bp (PE 150 bp).
The raw data were assembled using Mitoz 3.6 (Meng et al. 2019).We compared the sequences of the Tachinidae and manually annotated the sequence of V. turrita, and then the data were submitted to GenBank database through NCBI.The   (Katoh and Standley 2013) was used to align the 13 protein-coding genes (PCGs) and then trim them by trimAL (Capella-Guti� errez et al. 2009).Circular maps of the mitogenomes were prepared using Proksee (Grant et al. 2023) (Figure 2).

E-INSi method of MAFFT
To investigate the phylogenetic status of V. turrita, we used the mitogenome sequences of 23 species in the family Tachinidae, which included 17 sequences available for all subfamily Exoristinae in the GenBank database when we conducted this study.The mitogenome sequences of Lucilia sericata (Calliphoridae) and Sarcophaga crassipalpis (Sarcophagidae) were used as outgroups.The maximum-likelihood (ML) reconstruction was performed using IQ-TREE (Nguyen et al. 2015) with bootstrap set to 5000.The alignment, trimming, data concatenation, and tree building of sequences are all completed in PhyloSuite 1.2.2 (Zhang et al. 2020).

Results
The complete mitogenome of V. turrita (GenBank accession number: OR526344) is 17,387 bp in length.A total of 37 genes were annotated, consisting of 13 PCGs, two rRNA genes, 22 tRNA genes, and one non-coding region.The coverage depth map of V. turrita is detailed in Supplementary Figure S1.The nucleotide composition was 42.01% of A, 39.13% of T, 11.26% of C, 7.60% of G, and 81.14% of A þ T content.Most of the 13 PCGs used ATN as the start codon (ATG for CYTB, ND4, ND4L, COX2, COX3, and ATP6; ATT for ND1, ND2, ND5, and ND6; ATA for ND3 and ATC for ATP8), except that COX1 begins with codon TCG.The stop codon TAA is used to most of the PCGs (ND2, COX1, COX3, ATP8, ATP6, ND4L, CYTB, and ND6), whereas incomplete stop codon T is used by three PCGs (ND4, ND5, and COX2); ND3 and ND1 terminate with the codon TAG.
The results of the phylogenetic tree (Figure 3) show that the V. turrita belongs to the Exoristinae.The relationship between the genus Vibrissina and Compsilura is relatively close in the phylogenetic tree with 100% bootstrap support.The complete mitogenome of V. turrita will contribute to the in-depth research on molecular bases for the taxonomic system and phylogeny of Tachinidae.

Discussion and conclusions
The mitogenome sequences of V. turrita are similar in gene arrangements, gene number, and nucleotide composition with the other species in the Blondeliini, as a typical mitogenome.Because of the variation of length in the A þ T rich region, the length of mitochondrial genome is longer than the other reference species in the same tribe.
The phylogenetic results show that a relatively close relationship between Vibrissina and Compsilura, both belonging to the tribe Blondeliini in known classification systems (O'Hara et al. 2020).This result is consistent with the previous work of Stireman et al. (2019).Our works provided the phylogenetic information of Tachinidae at the mitochondrial genome level for inferring the phylogenetic relationship of Diptera species.

Figure 2 .
Figure2.Circular gene map of the V. turrita mitochondrial genome.The genes displayed inside and outside the circle are transcribed counterclockwise and clockwise, respectively.Purple represents tRNA genes, blue represents protein-coding genes, and green represents rRNA genes.